Natural and recombinant human ACE. In addition to soluble ACE, this kit recognizes membrane-bound forms from lysates of cultured cells and supernates.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Human ACE Immunoassay is a 4.5 hour solid phase ELISA designed to measure ACE in cell culture supernates, serum, plasma, and saliva. It contains NS0-expressed ectodomain of recombinant human sACE, and antibodies raised against the recombinant factor. Natural human ACE showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human ACE.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Saliva
Serum, Heparin Plasma
The recovery of ACE spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing high concentrations of ACE were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
ACE and ACE-2, two cell surface proteases, are important regulators of the renin-angiotensin system (RAS), which plays a key role in maintaining blood pressure homeostasis and fluid salt balance in mammals. Because of its location and specificity, ACE plays additional roles in immunity, reproduction and neuropeptide regulation. ACE exists in two isoforms. Somatic ACE comprises two highly similar protease domains whereas germinal ACE comprises a single protease domain. ACE-2 consists of a single protease domain and is an essential regulator of heart and lung function. It is also a cellular receptor for the Spike protein of SARS cornonavirus that causes severe acute respiratory syndrome.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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but also provides information about sample types, species, and experimental conditions.